Santos-Sanches Ilda, Chambel Lélia, Tenreiro Rogério
Faculdade de Ciências e Tecnologia (FCT), Centro de Recursos Microbiológicos (CREM) and Research Unit on Applied Molecular Biosciences (UCIBIO, REQUIMTE), Universidade Nova de Lisboa (UNL), Campus de Caparica, 2829-516, Caparica, Lisbon, Portugal,
Methods Mol Biol. 2015;1247:323-34. doi: 10.1007/978-1-4939-2004-4_23.
Pulsed-field gel electrophoresis (PFGE) separates large DNA molecules by the use of an alternating electrical field, such that greater size resolution can be obtained when compared to normal agarose gel electrophoresis. PFGE is often employed to track pathogens and is a valuable typing scheme to detect and differentiate strains. Particularly, the contour-clamped homogeneous electric field (CHEF) PFGE system is considered to be the gold standard for use in epidemiological studies of many bacterial pathogens. Here we describe a PFGE protocol that was applicable to the study of bovine streptococci, namely, Streptococcus agalactiae (group B Streptococcus, GBS), Streptococcus dysgalactiae subsp. dysgalactiae (group C Streptococcus, GCS), and Streptococcus uberis-which are relevant pathogens causing mastitis, a highly prevalent and costly disease in dairy industry due to antibiotherapy and loss in milk production.
脉冲场凝胶电泳(PFGE)通过使用交变电场来分离大的DNA分子,与普通琼脂糖凝胶电泳相比,这样能获得更高的大小分辨率。PFGE常用于追踪病原体,是检测和区分菌株的一种有价值的分型方法。特别是,轮廓夹钳均匀电场(CHEF)PFGE系统被认为是许多细菌病原体流行病学研究中的金标准。在此,我们描述一种适用于牛链球菌研究的PFGE方案,即无乳链球菌(B组链球菌,GBS)、停乳链球菌停乳亚种(C组链球菌,GCS)和乳房链球菌,它们是引起乳腺炎的相关病原体,乳腺炎是乳制品行业中一种因抗生素治疗和产奶量损失而高度流行且代价高昂的疾病。